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Gene delivery system having enhanced tumor-specific expression, and recombinant gene expression regulating sequence

a gene delivery system and tumor-specific technology, applied in the direction of dsdna viruses, drug compositions, biocide, etc., can solve the problems of limited blood supply in most tumor cells, limited persistence tumor treatment using viruses, etc., to improve tumor survival, improve tumor survival, and improve tumor survival

Active Publication Date: 2017-06-27
GENEMEDICINE CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Benefits of technology

The present inventors have developed a therapeutic agent with better tumor-specific gene delivery efficiency and increased oncolytic activity. They have found that using adenovirus with a specific expression regulatory sequence, including a HRE enhancer, an E2F promoter, and a TERT promoter, leads to significant improvements in oncolytic activity. This invention provides a gene delivery system with improved tumor-specific expression and a recombinant adenovirus with improved oncolytic activity.

Problems solved by technology

It turns out that persistent tumor treatments using virus are limited.
Blood supply in most of tumor cells is insufficient since tumor cells typically grow faster than endothelial cells generating blood vessels such that hypoxia is induced in tumor cells.

Method used

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  • Gene delivery system having enhanced tumor-specific expression, and recombinant gene expression regulating sequence
  • Gene delivery system having enhanced tumor-specific expression, and recombinant gene expression regulating sequence
  • Gene delivery system having enhanced tumor-specific expression, and recombinant gene expression regulating sequence

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examples

[0110]Preparation of Vector for E-Rd19, EE-Rd19, HE-Rd19 and HEE-Rd19

[0111]To construct adenovirus having an improved oncolytic activity by introducing 2 copies of E2F promoter, E2F promoter DNA in pSP72-E2F cloning vector was excised by SalI / NheI and inserted into XhoI / BamHI site of the pΔE1sp1B-E2F cloning vector to prepare pΔE1sp1B-E2F-E2F vector. Rb7Δ19 DNA digested by EcoRI / BamHI was subcloned into EcoRI / BglII of the pΔE1sp1B-E2F-E2F vector to prepare pΔE1sp1B-E2F-E2F-Rb7Δ19 adenovirus E1 shuttle vector having E2F promoter 2 copies. The prepared pΔE1sp1B-E2F-E2F-Rb7Δ19 shuttle vector was subjected to homologous recombination to E1 region of dl324 Bst as an adenovirus total vector, thereby preparing an oncolytic adenovirus EE-Rd19.

[0112]In addition, in order to develop oncolytic adenovirus having better activity in hypoxic tumors, HRE 6 copies and E2F promoter were combined to prepare oncolytic adenovirus regulated replication by the combined promoter. First, to construct E1 ade...

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Abstract

The present invention relates to a gene expression regulating sequence consisting of a combination of HRE, E2F and TERT, and to a gene delivery system having significantly improved selective tumor cell cytotoxicity using same, and more particularly, to a recombinant adenovirus. In addition, the present invention relates to a pharmaceutical antitumor composition comprising the recombinant adenovirus. The replication of the recombinant adenovirus of the present invention is tumor-specifically regulated by the novel gene expression regulating sequence of the present invention, thus enabling the recombinant adenovirus of the present invention to exhibit improved selective tumor cell cytotoxicity or apoptotic potential, and exhibit remarkably improved antitumor effects particularly in hypoxic conditions. In addition, the specific expression of the recombinant adenovirus in tumor cells may increase in vivo stability, and thus may induce greatly improved antitumor effects.

Description

CROSS REFERENCE TO RELATED APPLICATIONS AND CLAIM OF PRIORITY[0001]This patent application is a National Phase application under 35 U.S.C. §371 of International Application No. PCT / KR2011 / 004693, filed on Jun. 28, 2011, which claims priority to Korean Patent Application number 10-2010-0098498, filed on Oct. 8, 2010, entire contents of which are incorporated herein by reference.REFERENCE TO A SEQUENCE LISTING[0002]This application contains references to nucleic acid sequences which have been submitted concurrently herewith as the sequence listing text file “61652987_1.txt”, file size 2 KiloBytes (KB), created on 4 Jan. 2016. The aforementioned sequence listing is hereby incorporated by reference in its entirety pursuant to 37 C.F.R. §1.52(e)(5).FIELD OF THE INVENTION[0003]The present invention relates to a gene delivery system having a recombinant gene expression regulatory sequence with improved tumor-specific expression.DESCRIPTION OF THE RELATED ART[0004]A growth of malignant tumo...

Claims

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Application Information

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Patent Type & Authority Patents(United States)
IPC IPC(8): C12N15/86A61K35/761C12N15/85
CPCC12N15/85A61K35/761C12N15/86C12N2710/10332C12N2710/10343C12N2710/10345C12N2810/6018C12N2830/002C12N2830/008C12N2830/15C12N2830/85A61P35/00
Inventor YUN, CHAE OKYOON, A-RUM
Owner GENEMEDICINE CO LTD
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